SUWARSO SUWARSO
Balai Penelitian Tanaman Tembakau dan Serat Jl. Raya Karangploso, Kotak Pos 199, Malang

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ISOLASI FRAGMEN GEN PENYANDI PUTRESIN N-METILTRANSFERASE DAN QUINOLINAT FOSFORIBOSILTRANSFERASE ASAL TEMBAKAU LOKAL TEMANGGUNG (Nicotiana tabacum) SESANTI BASUKI; NURHAJATI AA MATTJIK; SUWARSO SUWARSO; DESTA WIRNAS; SUDARSONO SUDARSONO
Jurnal Penelitian Tanaman Industri Vol 17, No 3 (2011): September 2011
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jlittri.v17n3.2011.109-117

Abstract

ABSTRAKUpaya untuk menurunkan kandungan nikotin merupakan salah satuprioritas utama penelitian tembakau. Nikotin adalah senyawa alkaloidutama berpotensi dikonversi menjadi senyawa nor-nikotin yang bersifatkarsinogen. Gen PMT sebagai penyandi enzim putresin n-metiltransferase(PMT) dan gen QPT - penyandi enzim quinolinat fosforibosiltransferase(QPT) merupakan dua gen kunci yang berperan penting pada proses bio-sintesis nikotin. Penelitian ini bertujuan untuk mengisolasi potongan genPMT dan QPT asal tembakau lokal Indonesia, mengkarakterisasi danmenganalisis runutan DNA-nya. Tahapan penelitian dimulai dengan me-rancang primer degenerate berdasarkan informasi yang ada di pangkalandata Bank Gen NCBI (National Centre for Biotechnology Information),mengamplifikasi PCR menggunakan templat DNA genomik tembakaulokal cv. Sindoro1, mengklon potongan DNA hasil PCR dan menentukanrunutan DNA-nya. Hasil penelitian menunjukkan dari dua belas pasangprimer degenerate yang dirancang, hanya dua pasang primer yang meng-hasilkan potongan DNA hasil amplifikasi PCR, yaitu pasangan primerPMt-7 (F & R) untuk gen PMT dan primer QPt-3 (F & R) untuk gen QPT.Setelah dilakukan penentuan runutan DNA-nya, amplikon yang didapatdari hasil PCR dengan pasangan primer PMt-7 sebesar 1418 bp, sedangkanuntuk primer QPt-3 sebesar 205 bp. Runutan DNA gen PMT dan gen QPTasal tembakau lokal cv. Sindoro1 mempunyai tingkat kesamaan yang ting-gi dengan gen PMT dan gen QPT asal tembakau lainnya yang ada dipangkalan data Bank Gen NCBI.Kata kunci : Gen PMT, gen QPT, lintasan biosintesis nikotin, perunutanDNA, amplifikasi PCR, primer degenerateABSTRACTIsolation of Genes encoding Putrescine N-Methyl-transferase and Quinolinat Phosphoribosyl transferasederived from Temanggung Tobacco Cultivar (Nicotianatabacum)Reduction of nicotine content is one of the major objective intobacco research. Nicotine is the main alcaloid compound that potentiallycould be converted into a carcinogenic compound (nor-nicotine). The PMTgene encoding putrescine N-methyl transferase (PMT) and the QPT gene -encoding quinolinate phosphoribosyl transferase (QPT) are the two keyenzymes involved in nicotine biosynthesis. The objectives of this researchwere to isolate PMT and QPT gene fragments originated from Indonesianlocal tobacco, to characterize, and to analyze their DNA sequences. Theresearch activities included: degenerate primer design based oninformation available in the GenBank DNA Database NCBI (NationalCentre for Biotechnology Information), PCR amplification usingdegenerate primer and genomic DNA template of a local tobacco cv.Sindoro1, clone the PCR amplified products, and determine their DNAnucleotide sequences. Results of the experiment indicated that from 12degenerate primer pairs synthesized, only two were able to yield positivePCR amplified products. These primer pairs were PMt-7 (F & R primers)for PMT and QPt-3 (F & R primers) for QPT. After DNA sequencing, theamplified DNA product amplified using PMt-7 degenerate primer pairswere 1418 bp, while that using QPt-3 primer pairs were only 205 bp.Nucleotide sequences of PMT or QPT gene fragments originated fromlocal tobacco cv. Sindoro1 showed a high nucleotide sequences identity ascompared to that of the respective genes from other tobacco species thatwere available in the GenBank DNA Database NCBI.Key words: PMT gene, QPT gene, nicotine biosynthetic pathways, DNAsequencing, PCR amplification, degenerate primer
GALUR HARAPAN TEMBAKAU TEMANGGUNG PRODUKSI TINGGI DAN TAHAN PENYAKIT LINCAT FATKHUR ROCHMAN; SUWARSO SUWARSO; A.S. MURDIYATI A.S. MURDIYATI
Jurnal Penelitian Tanaman Industri Vol 13, No 2 (2007): JUNI 2007
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jlittri.v13n2.2007.57-63

Abstract

ABSTRAKMasalah  utama  tembakau  Temanggung  adalah  rendahnyaproduktivitas yang disebabkan oleh mundurnya daya dukung lahan karenaerosi dan endemi penyakit lincat (kompleks nematoda Meloidogyne spp,bakteri Ralstonia solanacearum, dan cendawan Phytophthora nicotianae).Saat ini telah diperoleh enam galur hasil persilangan dari varietas Sindoro1 (moderat tahan terhadap Ralstonia solanacearum tetapi rentan terhadapMeloidogyne spp. dan sangat rentan terhadap Phytophthora nicotianae)dengan tembakau virginia yang tahan terhadap ketiga patogen tersebut.Evaluasi terhadap hasil, indeks mutu, indeks tanaman dan ketahananterhadap ketiga patogen telah dilakukan di 3 lokasi selama tiga tahundengan rancangan acak kelompok tiga ulangan. Hasil penelitianmenunjukkan bahwa ada dua galur harapan yaitu (1) Galur A dengan rata-rata produktivitas : 0,880 ton rajangan kering per ha dan indeks mutu40,28 indeks tanaman 35,47 dan kadar nikotin 5,52%. Produktivitas galurA meningkat 48,08%, indeks mutu meningkat 4,87% dan indeks tanamanmeningkat 53,73% serta kadar nikotin menurun 15,06% dibanding varietasstandar. Galur A memiliki sifat moderat tahan terhadap bakteri Ralstoniasolanacearum dan toleran terhadap nematoda Meloidogyne spp. (2) GalurE dengan rata-rata produktivitas : 0,869 ton rajangan kering per ha, indeksmutu 36,01 indeks tanaman 31,87 dan kadar nikotin 6,00%. Produktivitasgalur E meningkat 46,23%, indeks mutu menurun 6,25% dan indekstanaman meningkat 38,12% serta kadar nikotin menurun 2,56% dibandingvarietas standar. Galur E memiliki sifat moderat tahan terhadap bakteriRalstonia solanacearum dan toleran terhadap nematoda Meloidogyne spp.Kata kunci : Tembakau,  Nicotiana  tabacum,  Temanggung, galurharapan, Ralstonia solanacearum, Meloidogyne spp,Phytophthora nicotianae, lahan lincat, Jawa TimurABSTRACTTemanggung tobacco promising lines with high producti-vity and resistant to lincat diseasedThe main problem in Temanggung tobacco cultivation is lowproductivity which is caused by increasing land erosion and invasion ofendemic disease called ‘lincat’. Lincat is a disease caused by a complexinvasion of three pathogens i.e. Meloidogyne spp. (root-knot nematode),Ralstonia solanacearum (bacteria) and Phytophthora nicotianae (fungus).Hybridization between variety Sindoro 1 (moderately resistant to R.solanacearum but susceptible to Meloidogyne spp. and highly susceptibleto P. nicotianae) and virginia tobacco (resistant to these three pathogens)has resulted in six lines. These lines were planted in three locations for 3years and were evaluated for their yield, grade index, crop index, andresistance intensity to these three pathogens, using randomized blockdesign with 3 replications. Two promising lines were resulted from thisevaluation i.e. (1) Line A, that has productivity of tobacco sliced : 0.880ton per hectare, grade index: 40.28 crop index: 35.47 and nicotine content:5.52%. In comparison to standard variety, the productivity, grade index,and crop index of this line increased of 48.08%, 4.87%, and 53.73%,respectively, and nicotine content decreased of 15.06%. Moreover, thisline is moderately resistant to R. solanacearum and tolerant toMeloidogyne spp. (2) Line E, that has productivity of tobacco sliced:0.869 ton per hectare, grade index: 36.01 crop index: 31.87 and nicotinecontent: 6.00%. This line also has increase productivity, and crop index of46.23% 38.12%, respectively, and has decrease grade index and nicotinecontent of 6.25% and 2.56%, respectively compared to standard variety.This line is moderately resistant to R. solanacearum and tolerant toMeloidogyne spp.Key words : Tobacco, Nicotiana tabacum, Temanggung, promising line,Ralstonia solanacearum, Meloidogyne spp., Phytophthoranicotianae, lincat land, East Jav
UJI PRODUKTIVITAS DAN MUTU TIGA VARIETAS TEMBAKAU ORIENTAL DI INDONESIA SUWARSO SUWARSO; SAMSURI TIRTOSASTRO; TITIEK YULIANTI; SUHARTO SUHARTO; SUSENO SUSENO; M. YASIN
Jurnal Penelitian Tanaman Industri Vol 16, No 3 (2010): September 2010
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jlittri.v16n3.2010.112-118

Abstract

ABSTRAKUntuk memperoleh varietas yang sesuai di Indonesia telahdilakukan pengujian tiga varietas tembakau oriental, yaitu Zichna, XanthiYaka dan Izmir di Desa Rejuno, Dero (Kabupaten Ngawi) danMargomulyo (Kabupaten Bojonegoro). Tipe tanah ketiga desa tersebutberturut-turut adalah lempung, liat, dan lempung berdebu. Pengujiandilakukan pada tiga periode tanam tahun 2007 dan 2008. Hasil penelitianmenunjukkan bahwa tipe tanah berpengaruh terhadap hasil dan indekstanaman, yang terbaik adalah tanah lempung seperti di Desa Rejuno.Berdasarkan hasil, indeks tanaman dan penilaian organoleptik, varietasyang terbaik adalah Zichna, hasilnya rata-rata 2,213 t/ha dan indekstanaman 91,66. Varietas tersebut menghasilkan sensasi iritasi dan impakrendah dan aroma sangat baik. Pada urutan berikutnya adalah varietasXanthi Yaka, hasilnya 1,742 t/ha, indeks tanaman 78,27. Berdasarkan hasilpenelitian tersebut maka kedua varietas tembakau oriental tersebut jugasesuai untuk rokok kretek di Indonesia. Keduanya sesuai ditanam pada tipetanah lempung atau tanah yang banyak mengandung kapur dan pasir.Kata kunci: Nicotiana tabacum L., tembakau oriental, varietas, IndonesiaABSTRACTProductivity and quality tests of three oriental tobacco varieties in IndonesiaThree oriental tobacco varieties were tested in villages of Rejuno,Dero (Ngawi regency), and Margomulyo (Bojonegoro regency) to find outsuitable variety to grow in Indonesia. Soil types of the three villages wereloam, clay, and silt loam, respectively. The tests were carried out for threeplanting series in 2007 and 2008. Research result showed that soil typesaffected several agronomic characters as well as yield and crop indices.The most suitable soil was silt loam as in Rejuno village. According toyield, crop index, and organoleptic evaluation, Zichna variety was the bestwith yield potential and crop index of 2.213 t/ha and 91.66.Organoleptically, the variety was low irritation and sensation impact andvery good in aroma. The second best variety was Xanthi Yaka with yieldpotential and crop index of 1.742 t/ha and 78.27. This research revealedthat Zichna and Xanthi Yaka varieties were suitable as raw material forclove cigarette. In addition, these two varieties were also well suited to begrown in Indonesia, especially on loamy and sandy soils with large amountof lime.Key words : Nicotiana tabacum L., oriental tobacco, variety, Indonesia
ISOLASI FRAGMEN GEN PENYANDI PUTRESIN N-METILTRANSFERASE DAN QUINOLINAT FOSFORIBOSILTRANSFERASE ASAL TEMBAKAU LOKAL TEMANGGUNG (Nicotiana tabacum) SESANTI BASUKI; NURHAJATI AA MATTJIK; SUWARSO SUWARSO; DESTA WIRNAS; SUDARSONO SUDARSONO
Jurnal Penelitian Tanaman Industri Vol 17, No 3 (2011): September 2011
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jlittri.v17n3.2011.109-117

Abstract

ABSTRAKUpaya untuk menurunkan kandungan nikotin merupakan salah satuprioritas utama penelitian tembakau. Nikotin adalah senyawa alkaloidutama berpotensi dikonversi menjadi senyawa nor-nikotin yang bersifatkarsinogen. Gen PMT sebagai penyandi enzim putresin n-metiltransferase(PMT) dan gen QPT - penyandi enzim quinolinat fosforibosiltransferase(QPT) merupakan dua gen kunci yang berperan penting pada proses bio-sintesis nikotin. Penelitian ini bertujuan untuk mengisolasi potongan genPMT dan QPT asal tembakau lokal Indonesia, mengkarakterisasi danmenganalisis runutan DNA-nya. Tahapan penelitian dimulai dengan me-rancang primer degenerate berdasarkan informasi yang ada di pangkalandata Bank Gen NCBI (National Centre for Biotechnology Information),mengamplifikasi PCR menggunakan templat DNA genomik tembakaulokal cv. Sindoro1, mengklon potongan DNA hasil PCR dan menentukanrunutan DNA-nya. Hasil penelitian menunjukkan dari dua belas pasangprimer degenerate yang dirancang, hanya dua pasang primer yang meng-hasilkan potongan DNA hasil amplifikasi PCR, yaitu pasangan primerPMt-7 (F & R) untuk gen PMT dan primer QPt-3 (F & R) untuk gen QPT.Setelah dilakukan penentuan runutan DNA-nya, amplikon yang didapatdari hasil PCR dengan pasangan primer PMt-7 sebesar 1418 bp, sedangkanuntuk primer QPt-3 sebesar 205 bp. Runutan DNA gen PMT dan gen QPTasal tembakau lokal cv. Sindoro1 mempunyai tingkat kesamaan yang ting-gi dengan gen PMT dan gen QPT asal tembakau lainnya yang ada dipangkalan data Bank Gen NCBI.Kata kunci : Gen PMT, gen QPT, lintasan biosintesis nikotin, perunutanDNA, amplifikasi PCR, primer degenerateABSTRACTIsolation of Genes encoding Putrescine N-Methyl-transferase and Quinolinat Phosphoribosyl transferasederived from Temanggung Tobacco Cultivar (Nicotianatabacum)Reduction of nicotine content is one of the major objective intobacco research. Nicotine is the main alcaloid compound that potentiallycould be converted into a carcinogenic compound (nor-nicotine). The PMTgene encoding putrescine N-methyl transferase (PMT) and the QPT gene -encoding quinolinate phosphoribosyl transferase (QPT) are the two keyenzymes involved in nicotine biosynthesis. The objectives of this researchwere to isolate PMT and QPT gene fragments originated from Indonesianlocal tobacco, to characterize, and to analyze their DNA sequences. Theresearch activities included: degenerate primer design based oninformation available in the GenBank DNA Database NCBI (NationalCentre for Biotechnology Information), PCR amplification usingdegenerate primer and genomic DNA template of a local tobacco cv.Sindoro1, clone the PCR amplified products, and determine their DNAnucleotide sequences. Results of the experiment indicated that from 12degenerate primer pairs synthesized, only two were able to yield positivePCR amplified products. These primer pairs were PMt-7 (F & R primers)for PMT and QPt-3 (F & R primers) for QPT. After DNA sequencing, theamplified DNA product amplified using PMt-7 degenerate primer pairswere 1418 bp, while that using QPt-3 primer pairs were only 205 bp.Nucleotide sequences of PMT or QPT gene fragments originated fromlocal tobacco cv. Sindoro1 showed a high nucleotide sequences identity ascompared to that of the respective genes from other tobacco species thatwere available in the GenBank DNA Database NCBI.Key words: PMT gene, QPT gene, nicotine biosynthetic pathways, DNAsequencing, PCR amplification, degenerate primer
UJI PRODUKTIVITAS DAN MUTU TIGA VARIETAS TEMBAKAU ORIENTAL DI INDONESIA SUWARSO SUWARSO; SAMSURI TIRTOSASTRO; TITIEK YULIANTI; SUHARTO SUHARTO; SUSENO SUSENO; M. YASIN
Jurnal Penelitian Tanaman Industri Vol 16, No 3 (2010): September 2010
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jlittri.v16n3.2010.112-118

Abstract

ABSTRAKUntuk memperoleh varietas yang sesuai di Indonesia telahdilakukan pengujian tiga varietas tembakau oriental, yaitu Zichna, XanthiYaka dan Izmir di Desa Rejuno, Dero (Kabupaten Ngawi) danMargomulyo (Kabupaten Bojonegoro). Tipe tanah ketiga desa tersebutberturut-turut adalah lempung, liat, dan lempung berdebu. Pengujiandilakukan pada tiga periode tanam tahun 2007 dan 2008. Hasil penelitianmenunjukkan bahwa tipe tanah berpengaruh terhadap hasil dan indekstanaman, yang terbaik adalah tanah lempung seperti di Desa Rejuno.Berdasarkan hasil, indeks tanaman dan penilaian organoleptik, varietasyang terbaik adalah Zichna, hasilnya rata-rata 2,213 t/ha dan indekstanaman 91,66. Varietas tersebut menghasilkan sensasi iritasi dan impakrendah dan aroma sangat baik. Pada urutan berikutnya adalah varietasXanthi Yaka, hasilnya 1,742 t/ha, indeks tanaman 78,27. Berdasarkan hasilpenelitian tersebut maka kedua varietas tembakau oriental tersebut jugasesuai untuk rokok kretek di Indonesia. Keduanya sesuai ditanam pada tipetanah lempung atau tanah yang banyak mengandung kapur dan pasir.Kata kunci: Nicotiana tabacum L., tembakau oriental, varietas, IndonesiaABSTRACTProductivity and quality tests of three oriental tobacco varieties in IndonesiaThree oriental tobacco varieties were tested in villages of Rejuno,Dero (Ngawi regency), and Margomulyo (Bojonegoro regency) to find outsuitable variety to grow in Indonesia. Soil types of the three villages wereloam, clay, and silt loam, respectively. The tests were carried out for threeplanting series in 2007 and 2008. Research result showed that soil typesaffected several agronomic characters as well as yield and crop indices.The most suitable soil was silt loam as in Rejuno village. According toyield, crop index, and organoleptic evaluation, Zichna variety was the bestwith yield potential and crop index of 2.213 t/ha and 91.66.Organoleptically, the variety was low irritation and sensation impact andvery good in aroma. The second best variety was Xanthi Yaka with yieldpotential and crop index of 1.742 t/ha and 78.27. This research revealedthat Zichna and Xanthi Yaka varieties were suitable as raw material forclove cigarette. In addition, these two varieties were also well suited to begrown in Indonesia, especially on loamy and sandy soils with large amountof lime.Key words : Nicotiana tabacum L., oriental tobacco, variety, Indonesia